SERS immunoassay based on the capture and concentration of antigen-assembled gold nanoparticles

Lopez, Arielle; Lovato, Francis L.; Oh, Soo Hee; Lai, Yen H.; Filbrun, Seth L.; Driskell, Elizabeth A.; Driskell, Jeremy D.

doi:10.1016/j.talanta.2015.08.065

Cited by 51 publications

(42 citation statements)

References 36 publications

(41 reference statements)

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“…Throughout the B. burgdorferi infection, there are few circulating organisms, and the organism burden in affected organs is low. Concentration methods can be categorized as follows: removal of solute or water [16], electrophoretic or chromatographic separation [17], and affinity capture [18][19][20]. Along with target enrichment, these methods may require exclusion of masking proteins such as common abundant proteins (frequently a preparative step prior to mass spectrometry detection of proteins).…”

Section: Detection Of Bacterial Antigensmentioning

confidence: 99%

Direct Diagnostic Tests for Lyme Disease

Schutzer

Body²,

Boyle

et al. 2018

Clinical Infectious Diseases

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Borrelia burgdorferi was discovered to be the cause of Lyme disease in 1983, leading to seroassays. The 1994 serodiagnostic testing guidelines predated a full understanding of key B. burgdorferi antigens and have a number of shortcomings. These serologic tests cannot distinguish active infection, past infection, or reinfection. Reliable direct-detection methods for active B. burgdorferi infection have been lacking in the past but are needed and appear achievable. New approaches have effectively been applied to other emerging infections and show promise in direct detection of B. burgdorferi infections.

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Section: Detection Of Bacterial Antigensmentioning

confidence: 99%

Direct Diagnostic Tests for Lyme Disease

Schutzer

Body²,

Boyle

et al. 2018

Clinical Infectious Diseases

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“…prepared the gold nanoparticles with caffeic acid as reducer, and the particle geometry could be controlled effectively by changing the caffeic acid concentration. Although silver nanoparticles commonly had higher SERS activity than gold nanoparticles, its stability was less than gold nanoparticles, and it was easier to be oxidized678910. Therefore, the preparation of gold nanoparticle with high SERS activity was very important to SERS quantitative analysis.…”

mentioning

confidence: 99%

“…Therefore, the preparation of gold nanoparticle with high SERS activity was very important to SERS quantitative analysis. Lopez et al 6. developed a simple and sensitive SERS detection immunoassay based on antigen-mediated aggregation of gold nanoparticles with a detection limit of 1.9 ng/mL, and it was 20-fold higher than the ELISA assay.…”

mentioning

confidence: 99%

SERS spectral study of HAuCl4-cysteine nanocatalytic reaction and its application for detection of heparin sodium with label-free VB4r molecular probe

Wang

Jiang

Qin

et al. 2017

Sci Rep

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In the presence of nanocatalyst, L-cysteine reduce HAuCl4 rapidly to form gold nanoparticles (AuNP), and a quick nanocatalytic preparation procedure was established for Au/AuNP sol with highly active surface enhanced Raman scattering (SERS) effect and good stability. The nanoreaction was also studied by absorption, resonance Rayleigh scattering (RRS), transmission electron microscopy (TEM) and energy spectra. In the selected conditions, the analyte heparin sodium (HS) could react with victoria blue 4 R (VB4r) to form associated complexes which have very weak SERS effect to make the SERS signals decrease. The SERS signals at 1617 cm−1 reduced linearly with HS concentration increasing. Upon addition of FeCl3, it hydrolyzed to form stable Fe(OH)3 sol platform that carried SERS active Au/AuNPs to enhance the sensitivity. Accordingly, we established a SERS quantitative analysis method in the sol substrate of Fe(OH)3-Au/AuNPs, with a linear range of 0.5–75 ng/mL HS and a detection limit of 0.2 ng/mL. HS in real samples was determined, with a relative standard deviation of 2.65–7.63% and a recovery of 99.3–101%.

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“…[6][7][8][9] Surface enhanced Raman spectroscopy (SERS) is a widely known technique in which modified gold nanoparticles are used for the detection of many analytess at once. [10][11][12][13] Many diagnostic techniques employing metallic nanoparticles for the detection of specific biomarkers entail proteins to be adsorbed onto the surface of the nanoparticle due to the high specificity and selectivity of proteins. Antibody-antigen interactions are highly exploited in many of these detection techniques.…”

Section: Gold Nanoparticles (Aunp) For In-vitro Diagnosticsmentioning

confidence: 99%

“…Antibody-antigen interactions are highly exploited in many of these detection techniques. 11 Consequently, conjugation is fundamental to the stability, functionality, and overall success of AuNP based detection platforms. Similarly, efficiency of the conjugation chemistry, i.e., the number of adsorbed proteins, is imperative to the optimization of novel immunosensors.…”

Section: Gold Nanoparticles (Aunp) For In-vitro Diagnosticsmentioning

confidence: 99%

Formation, Characterization, And Optimization Of Antibody-Gold Nanoparticle Conjugates

Filbrun¹

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Protein modified gold nanoparticle based immunoassays are the basis of many novel detection techniques. There are many groups working on novel immunoassays but there is still much to understand about how many proteins are attached onto each nanoparticle and much to improve on immobilization methods. This thesis work is devoted to improving techniques for the quantification of protein immobilized on the gold nanoparticle and development of a novel approach to immobilize protein independent of pH.The ability to evaluate antibody immobilization onto gold nanoparticles is critical for assessing coupling chemistry and optimizing the sensitivity of nanoparticle-enabled biosensors.Herein, we developed a fluorescence-based method for directly quantifying antibodies bound onto gold nanoparticles. Antibody-modified gold nanoparticles were treated with KI/I2 etchant to dissolve the gold nanoparticles. A desalting spin column was used to recover the antibody released from the nanoparticles, and NanoOrange, a fluorescent dye, was used to quantify the antibody. We determined 309 ± 93 antibodies adsorb onto each 60 nm gold nanoparticle (2.6 × 10 10 NP/mL), which is consistent with a fully adsorbed monolayer based on the footprint of an IgG molecule. Moreover, the increase in hydrodynamic diameter of the conjugated nanoparticle (76 nm) compared to that of the unconjugated nanoparticle (62 nm) confirmed that multilayers did not form. A more conventional method of indirectly quantifying the adsorbed antibody by analysis of the supernatant overestimated the antibody surface coverage (660 ± 87 antibodies per nanoparticle); thus, we propose the method described herein as a more accurate alternative to the conventional approach.The immobilization of antibody onto gold nanoparticles is important for many novel nanoparticle based immunoassays. Current methods of immobilization are limited by the inability to immobilize antibody onto gold nanoparticles over a range of pH values. Direct adsorption requires the pH to be slightly higher than the isoelectric point of the antibody and covalent attachment via bifunctional crosslinking chemistry molecules requires a specific pH as well. This is an issue when working with multiple antibodies at once for multiplex detection. In this thesis, we present a new method of immobilization by which the antibody is modified via a molecule with a N-hydroxysuccinimidyl ester group, then adsorbed onto a gold nanoparticle. We demonstrate that modification of antibodies allows for adsorption onto gold nanoparticles independent of pH. Furthermore, we show that this modification method is applicable to multiple antibodies. Finally, we show that these modified antibodies are active and comparable to conventional assays.

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SERS immunoassay based on the capture and concentration of antigen-assembled gold nanoparticles

Cited by 51 publications

References 36 publications

Direct Diagnostic Tests for Lyme Disease

Direct Diagnostic Tests for Lyme Disease

SERS spectral study of HAuCl4-cysteine nanocatalytic reaction and its application for detection of heparin sodium with label-free VB4r molecular probe

Formation, Characterization, And Optimization Of Antibody-Gold Nanoparticle Conjugates

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